Hela cells jcrb9004

HeLa cells (JCRB9004) and Huh7 cells (JCRB0403) were obtained from the Japanese Collection of Research Bioresources (Osaka, Japan) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Japan) supplemented with 10% fetal bovine serum (SAFC, MO, USA). The DFO-resistant cell lines were established by gradually increasing the DFO concentration (started from 3 μM) over a duration of approximately 6 months. Cells (2000~6000/well) were seeded in 96-well plates and treated with different concentrations (0~100 μM) of DFO, and cell proliferation was determined by measuring the area of the cells using the Incucyte HD imaging system (Essen BioScience, Ann Arbor, MI). The combination activity of the drugs was estimated with the CalcuSyn software program (Biosoft, Ferguson, MO). Briefly, this program determines the combination index (CI), a quantitative measure of the degree of drug interactions. The information related to drugs is provided in Supplemental Table 1.